Physiological adaptation of the Asian corn borer Ostrinia furnacalis to chemical defenses of its host plant, maize

A number of gramineous plants such as maize contain cyclic hydroxamic acids (cHx) that are toxic to many herbivores. Among the Ostrinia species found in Japan, the Asian corn borer Ostrinia furnacalis is the only one that utilizes maize, a gramineous plant. We used O. furnacalis and two congeners, Ostrinia scapulalis and Ostrinia latipennis, to obtain insights into the physiological adaptation of O. furnacalis to cHx. When an artificial diet containing a low concentration (0.3 mg/g diet) of cHx was fed to the larvae of O. furnacalis and O. scapulalis, larval growth and survival were significantly less affected in O. furnacalis than O. scapulalis. An artificial diet containing a high level (0.7 mg/g diet) of cHx was found to severely retard the growth of both species, albeit to different degrees. In an assay in vitro, homogenate of the digestive tract of O. furnacalis larvae degraded cHx more rapidly than that of O. scapulalis or O. latipennis. The degradation was found to be enzymatic and dependent on a cofactor, UDP-glucose, suggesting that UDP-glucosyltransferase or other UDP-glucose-dependent enzymes were involved. This enzymatic adaptation probably has enabled O. furnacalis to utilize plants containing cHx.     

A male-specific odorant receptor conserved through the evolution of sex pheromones in Ostrinia moth species

In many moths, mate-finding communication is mediated by the female sex pheromones. Since differentiation of sex pheromones is often associated with speciation, it is intriguing to know how the changes in female sex pheromone have been tracked by the pheromone recognition system of the males. A male-specific odorant receptor was found to have been conserved through the evolution of sex pheromone communication systems in the genus Ostrinia (Lepidoptera: Crambidae). In an effort to characterize pheromone receptors of O. scapulalis, which uses a mixture of (E)-11- and (Z)-11-tetradecenyl acetates as a sex pheromone, we cloned a gene (OscaOR1) encoding a male-specific odorant receptor. In addition, we cloned a gene of the Or83b family (OscaOR2). Functional assays using Xenopus oocytes co-expressing OscaOR1 and OscaOR2 have shown that OscaOR1 is, unexpectedly, a receptor of (E)-11-tetradecenol (E11-14:OH), a single pheromone component of a congener O. latipennis. Subsequent studies on O. latipennis showed that this species indeed has a gene orthologous to OscaOR1 (OlatOR1), a functional assay of which confirmed it to be a gene encoding the receptor of E11-14:OH. Furthermore, investigations of six other Ostrinia species have revealed that all of them have a gene orthologous to OscaOR1, although none of these species, except O. ovalipennis, a species most closely related to O. latipennis, uses E11-14:OH as the pheromone component. The present findings suggest that the male-specific receptor of E11-14:OH was acquired before the divergence of the genus Ostrinia, and functionally retained through the evolution of this genus.     

Sex pheromone of Ostrinia sp. newly found on the leopard plant Farfugium japonicum

Recently, larvae of Ostrinia were found feeding on the leopard plant Farfugium japonicum (Asteraceae), previously unrecorded as a host plant of this genus. The adult moths that developed from these borers were morphologically similar to, but distinct from, Ostrinia zaguliaevi, a monophagous species specialized for feeding on another Asteraceae plant, the butterbur Petasites japonicus. Although the taxonomical status of the moth feeding on F. japonicum is to be determined, distinct morphological differences in the adults strongly suggest this to be a new species (hereafter referred to as O. sp.). To gain an insight into the reproductive isolation between O. sp. and other members of the genus Ostrinia, the female sex pheromone and the males’ response to it were investigated using samples collected from F. japonicum. (Z)‐9‐tetradecenyl acetate (Z9‐14:OAc), (Z)‐11‐tetradecenyl acetate (Z11‐14:OAc), (E)‐11‐tetradecenyl acetate (E11‐14:OAc), tetradecyl acetate, and (Z)‐11‐hexadecenyl acetate were identified as candidates for sex pheromone components by analyses using gas chromatographs coupled to a mass spectrometer (GC‐MS) and electroantennographic detector (GC‐EAD). A series of bioassays of male responses in a wind‐tunnel and a field cage indicated that the former three compounds are essential for attracting males, and the latter two have no synergistic effect on the attraction. We therefore concluded that Z9‐14:OAc, Z11‐14:OAc and E11‐14:OAc are the sex pheromone components of O. sp. Although the same three compounds are used as the sex pheromone components of O. zaguliaevi and another congener, Ostrinia zealis, the blend proportions differed greatly among the three (Z9‐14:OAc/Z11‐14:OAc/E11‐14:OAc = 18/76/6 in O. sp., 45/50/5 in O. zaguliaevi and 70/6/24 in O. zealis). Differences in sex pheromones could contribute to the reproductive isolation between O. sp. and the other two Ostrinia species if males of each species exhibit a narrow window of response to their own blend ratio.     

Study of the genetics of female sex pheromone production and male behavioral response in a moth, Ostrinia orientalis

Inheritance patterns of female sex pheromone production and male behavioral response were studied in Ostrinia orientalis. Results showed that the production of the female sex pheromone in O. orientalis was mainly controlled by a single autosomal factor, while the male behavioral response was controlled by a sex‐linked major gene.     

Did the introduction of maize into Europe provide enemy‐free space to Ostrinia nubilalis? Parasitism differences between two sibling species of the genus Ostrinia

We examined whether maize offers enemy‐free space (EFS) to its pest Ostrinia nubilalis, and may thereby have contributed to its divergence from the sibling species, Ostrinia scapulalis, feeding mainly on mugwort, when introduced into Europe five centuries ago. We collected Ostrinia larvae on maize (70 populations, 8425 individuals) and mugwort (10 populations, 1184 individuals) and recorded parasitism using both traditional (counting emerging parasitoids) and molecular methods (detection by specific polymerase chain reaction). The main parasitoid was Macrocentrus cingulum (Braconidae). On mugwort, parasitism was twice that on maize, and parasitoid‐related mortality was 8 times higher. This suggests that maize affords substantial EFS to Ostrinia feeding on it. The lower Mortality:Infestation ratio in maize suggests that O. nubilalis’ immune response might be stronger than that of O. scapulalis. If so, adapting to maize and diverging from O. scapulalis would decrease the impact of parasitism on O. nubilalis at both ecological and evolutionary levels.     

Wolbachia infection and an all-female trait in Ostrinia orientalis and Ostrinia zaguliaevi

Sex ratio distortion toward females (SR trait), induced by a Wolbachia bacterium, has been reported in two species of the Ostrinia furnacalis group, viz., O. furnacalis, and O. scapulalis (Lepidoptera: Crambidae). In addition, an SR trait caused by abacterial, unidentified agent(s) is known in O. scapulalis. Here we examined the SR trait in four other species of the furnacalis group, viz., O. orientalis, O. zaguliaevi, and O. zealis from Japan, and O. nubilalis from central and eastern Europe. Wolbachia infection was detected in three O. orientalis females and in one O. zaguliaevi female, and the infection was always associated with the SR trait. In terms of wsp and ftsZ gene sequences, Wolbachia strains in O. orientalis and O. zaguliaevi were indistinguishable from each other, or from those in O. furnacalis and O. scapulalis. These findings suggest that Wolbachia strains in the four Ostrinia species are probably identical. In addition, one O. zealis female, which was negative in diagnostic PCRs for Wolbachia and general bacteria, produced an all‐female brood. This trait was very similar to the SR trait caused by abacterial agent(s) in O. scapulalis.     

General biology of Ostrinia palustralis memnialis (Walker), a potential biological control agent of Rumex spp. in Korea

Ostrinia palustralis memnialis is an oligophagous insect that feeds on the species of the genus Rumex. Continuous rearing of O. palustralis memnialis was carried out from 2004 to 2005, and its general biology was studied. The preliminary host specificity test showed a promising result for O. palustralis memnialis as a potential biological control agent for Rumex spp.     

Reconsidering the taxomony of several Ostrinia species in the light of reproductive isolation: a tale for Ernst Mayr

We reconsider the taxonomy of a group of closely related Ostrinia spp., illustrating how useful Mayr's biological species concept remains for studying speciation patterns and processes. We review and re-analyse recent data on Ostrinia scapulalis , Ostrinia nubilalis , Ostrinia narynensis and Ostrinia orientalis , along with those obtained over > 45 years in the former Soviet Union. The ten species of the 'trilobed uncus' group in the Ostrinia genus are classified into subgroups according to male mid-tibia morphology. However, none of the characters that further discriminate between them (female sex pheromones, male genitalia and calling time) varies together with male mid-tibia morphology, and neither do molecular markers. Moreover, male mid-tibia morphology appears to depend on only two diallelic loci and seems to be unrelated to reproductive isolation between Ostrinia taxa. By contrast, reproductive isolation is strongly related to host-plant type. In accordance with Mayr's species concept, we thus propose a revision of the trilobed uncus Ostrinia spp. based primarily on host-plant type. We propose that O. narynensis Mutuura & Munroe, 1970 ( syn. nov. ) and O. orientalis Mutuura & Munroe, 1970 ( syn. nov. ) be synonymized with O. scapulalis (Walker, 1859). We further demonstrate that O. nubilalis auctt. pro parte feeding on mugwort, hop, and several other dicotyledons (previously called the ' O. nubilalis mugwort-race' in France) also belongs to O. scapulalis . Consequently, we propose that only O. nubilalis specimens feeding on maize (the former French ' O. nubilalis maize-race') belong to O. nubilalis (Hübner, 1796). The implications of this revision are discussed.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 49–72.     

Ostrinia spp. in Japan: their host plants and sex pheromones

To contribute to the understanding of the genus Ostrinia (Lepidoptera; Pyralidae) in Japan, we collected larvae of Ostrinia spp. from known host plants and plants not recorded as hosts, and we examined the morphology and sex pheromones of the adults obtained. Consequently, the host plant ranges of the 7 Ostrinia spp. in Japan were clarified, and the sex pheromones of the 5 species O. scapulalis, O. zealis, O. zaguliaevi, O. palustralis and O. latipennis were identified in addition to that of the Asian corn borer O. furnacalis. The phylogenetic relationships of Japanese Ostrinia spp., with reference to the European corn borer O. nubilalis, are discussed based on these findings and results of molecular phylogenetic analyses.     

亚洲玉米螟与欧洲玉米螟混生区的研究

生殖隔离试验、形态鉴定、网室内性信息素活性反应和食性试验的结果表明 :张家口苍耳中的玉米螟与新疆伊宁的欧洲玉米螟同种 ,而为害玉米的优势种与广东阳山的亚洲玉米螟同种。张家口为害玉米的种群中 ,有部分雄蛾对欧洲玉米螟性信息素及苍耳中的雌处女蛾具有反应。苍耳和玉米中的种群在玉米、谷子、高梁、苍耳、草和大马蓼等 6种寄主上均可产卵和取食为害 ,并能正常完成生活史。研究结果证实张家口为亚洲玉米螟和欧洲玉米螟混生区 ,而且在玉米中很可能有少量欧洲玉米螟与亚洲玉米螟混生。     

Multiple Δ11-desaturase genes selectively used for sex pheromone biosynthesis are conserved in Ostrinia moth genomes

Regulation of the expression of fatty acyl-CoA desaturases, which introduce a double bond into the fatty acid moiety of the substrate, is crucial for the production of species-specific sex pheromones in moths. In Ostrinia moths, two distinct Δ11-desaturases and a Δ14-desaturase are known to be selectively used in the biosynthesis of sex pheromones. Of the two Δ11-desaturases, one identified from Ostrinia nubilalis and Ostrinia scapulalis, Z/EΔ11, forms the Z and E isomers of a double bond at position 11, whereas the other identified from Ostrinia latipennis, LATPG1(=EΔ11), exclusively forms an E double bond at position 11. Since the retroposon(ezi)-fused, non-functional Δ11-desaturase gene, ezi-Δ11α, in the genomes of O. nubilalis and O. furnacalis was previously suggested to be an orthologue of latpg1, we here explored Z/EΔ11 orthologues in the genome of O. latipennis. We newly identified two Δ11-desaturase genes, latpg2 and latpg3, which were orthologous to ezi-Δ11β and Z/EΔ11, respectively. We found that an ezi-like element was integrated in intron 1 of latpg1, and confirmed that only latpg1 was expressed in the pheromone gland of O. latipennis. Thus, at least three Δ11-desaturase genes are present in the genome of O. latipennis, and latpg1 is selectively transcribed in the pheromone gland of this moth. The non-functionality of ezi-inserted desaturase genes in O. nubilalis and O. furnacalis may not be a direct consequence of the insertion of an ezi- or ezi-like element into the gene.     

Seed inoculation with beneficial rhizobacteria affects European corn borer (Lepidoptera: Pyralidae) oviposition on maize plants

Larvae of Ostrinia nubilalis (Hübner) cause significant damage to maize ears and reduce market value of fresh sweet corn. Females rely on volatile cues to locate and oviposit preferentially on maize plants. In addition, oviposition behavior of females is influenced by soil management practices as they usually lay more eggs on maize plants grown on conventional soil than on organic soils that harbor rich microbial diversity. Since some plant growth‐promoting rhizobacteria (PGPR) are known to mediate plant health via suppression of soil pathogens and enhanced uptake of nutrients; we hypothesized that inoculation of maize seeds with PGPR will alter emission of maize volatile and reduce the attractiveness of plants to ovipositing O. nubilalis. Plants treated with the single PGPR strain Bacillus pumilus INR‐7, two PGPR mixtures (Blend‐8 or Blend‐9) or untreated plants were presented to O. nubilalis females in oviposition choice bioassays. Headspace volatile organic compounds (VOCs) from the plants were analyzed by gas chromatography–mass spectrometry (GC–MS). Ostrinia nubilalis laid significantly fewer eggs on PGPR‐treated plants compared to untreated plants. In two‐choice oviposition experiments, significantly higher numbers of eggs were laid on untreated plants compared to PGPR‐treated plants. PGPR‐treated plants emitted fewer VOCs than untreated plants which, in part, explains the relatively fewer eggs on PGPR‐treated plants. These results indicate that selected PGPR treatments can alter maize plant volatiles with important ramifications for plant‐insect interactions. The implication of this finding is discussed in the context of integrated management of soil health to improve crop resistance to biotic stressors.     

Age‐dependent changes in tolerance to cold and accumulation of cryoprotectants in overwintering and non‐overwintering larvae of European corn borer Ostrinia nubilalis

Abstract The age‐dependent cold hardiness profile of Ostrinia nubilalis is compared between nondiapausing and diapausing larvae, as well as with field‐collected larvae. The results suggest that both cold tolerance and accumulation of cryoprotectants depends upon the age of O. nubilalis larva. Late fifth‐instar nondiapausing larvae are more cold tolerant than younger fifth‐instars because they show enhanced ability to withstand sub‐zero temperatures. No appreciable difference is observed between the experimental groups of diapausing larvae as far as their supercooling ability and tolerance at sub‐zero temperatures above the supercooling point. In general, both field‐collected and diapausing larvae are more cold tolerant than nondiapausing larvae, indicating a direct link between diapause and cold hardiness. The age of diapausing larvae affects the ability to accumulate glycerol. Glycerol levels of 45‐day‐old diapausing larvae are significantly higher (2.7‐fold) compared with 90‐day‐old diapausing larvae. Moreover, diapausing larvae display a five‐ to 13‐fold higher glycerol content compared with nondiapausing larvae. There is a trend for an age‐dependent cold hardiness profile in O. nubilalis and further tests that could demonstrate a causal relationship between age and cold tolerance are needed.     

Genetic analysis and population survey of sex pheromone variation in the adzuki bean borer moth, Ostrinia scapulalis

Sexual communication in many moths occurs between females emitting a sex pheromone and males responding to it. Females of Ostrinia scapulalis (Lepidoptera: Crambidae) show a large variation in blend ratios of the two sex pheromone components (E)‐ and (Z)‐11‐tetradecenyl acetates. E type females produce a pheromone with a high percentage of (E)‐11‐tetradecenyl acetate, whereas Z type females produce the opposite blend. We established laboratory cultures of E and Z types. Females of the F₁ generation produced an intermediate blend (I type) in both reciprocal crosses of the E and Z cultures. Results of further crossing experiments suggested that the three pheromone types are primarily controlled by a single autosomal locus with two alleles. Also, analyses of the variation in pheromone blend within F₁, backcross and F₂ families suggested that other genetic factors modify the pheromone blend of the I and Z types. Investigation of the pheromone variation in natural populations at 14 localities in Japan has shown that the E type was predominant in northern Japan, whereas the pheromone was highly polymorphic in central Japan. At a locality in central Japan, the pheromone was constantly polymorphic for several years, and the pheromone type frequencies did not deviate from Hardy–Weinberg expectations, providing no evidence of selection or assortative mating between the pheromone types. Analyses of pheromone variation within families derived from feral females indicated that matings between a pair with different genotypes for pheromone production was occurring in natural populations. Overall, this study showed that the genetic basis of the pheromone variation in O. scapulalis is very similar to that in its sibling species Ostrinia nubilalis although the state of pheromone polymorphisms in natural populations appears to differ between the two species. © 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84 , 143–160.     

Genetic structure of European and Mediterranean maize borer populations on several wild and cultivated host plants

Target pests may become resistant to Bacillus thuringiensis (Bt) toxins produced by trangenic maize (Zea mays L.). Untreated refuge areas are set aside to conserve high frequencies of susceptibility alleles: a delay in resistance evolution is expected if susceptible individuals from refuges mate randomly with resistant individuals from Bt fields. In principle, refuges can be toxin‐free maize or any other plant, provided it hosts sufficiently large pest populations mating randomly with populations from Bt‐maize fields. Our aim was to examine the suitability of several cultivated or weedy plants [pepper (Capsicum frutescens L.), sorghum (Sorghum spec.), sunflower (Helianthus annuus L.), cocklebur (Xanthium spec.), cantaloupe (Cucumis melo L.), and hop (Humulus lupulus L.)] as refuges for Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) and Sesamia nonagrioides Lefebvre (Lepidoptera: Noctuidae), two major maize pests in southern Europe. Larvae of both species were collected on these plants. Their genetic population structure was examined at several allozyme loci. We found little or no evidence for an influence of geographic distance, but detected a significant host‐plant effect on the genetic differentiation for both species. Ostrinia nubilalis populations from sunflower, pepper, cocklebur, and sorghum appear to belong to the same genetic entity as populations collected on maize, but to differ from populations on hop. Accordingly, females from pepper and cocklebur produced exclusively the ‘Z’ type sexual pheromone, which, in France, characterizes populations developing on maize. Qualitatively, these plants (except hop) could thus serve as refuges for O. nubilalis; however, they may be of little use quantitatively as they were found much less infested than maize. Sesamia nonagrioides populations on maize and sorghum reached comparable densities, but a slight genetic differentiation was detected between both. The degree of assortative mating between populations feeding on both hosts must therefore be assessed before sorghum can be considered as a suitable refuge for this species.     

Scanning the European corn borer (Ostrinia spp.) genome for adaptive divergence between host-affiliated sibling species

It has recently been shown that the European corn borer, a major pest of maize crops, is actually composed of two genetically differentiated and reproductively isolated taxa, which are found in sympatry over a wide geographical range in Eurasia. Each taxon is adapted to specific host plants: Ostrinia nubilalis feeds mainly on maize, while O. scapulalis feeds mainly on hop or mugwort. Here, we present a genome scan approach as a first step towards an integrated molecular analysis of the adaptive genomic divergence between O. nubilalis and O. scapulalis. We analysed 609 AFLP marker loci in replicate samples of sympatric populations of Ostrinia spp. collected on maize, hop and mugwort, in France. Using two genome scan methods based on the analysis of population differentiation, we found a set of 28 outlier loci that departed from the neutral expectation in one or the other method (of which a subset of 14 loci were common to both methods), which showed a significantly increased differentiation between O. nubilalis and O. scapulalis, when compared to the rest of the genome. A subset of 12 outlier loci were sequenced, of which 7 were successfully re‐amplified as target candidate loci. Three of these showed homology with annotated lepidopteran sequences from public nucleotide databases.     

Species‐specific elicitors induce tea leaf to arrest the endoparasitoid Ascogaster reticulata (Hymenoptera: Braconidae)

Ascogaster reticulata Watanabe (Hymenoptera: Braconidae) is an egg‐larval endoparasitoid of the smaller tea tortrix, Adoxophyes honmai Yasuda. Recent studies have examined tritrophic interactions among Camellia sinensis, A. honmai and A. reticulata, but the effect of non‐host insects on the induction of tea plant that may affect foraging behaviour of A. reticulata remains unclear. In this study, we selected two non‐host insects, Homona magnanima Diakonoff and Ostrinia furnacalis (Guenée), as representative species in our bioassays. Tea leaves were treated with homogenized female reproductive tissues of a non‐host insect in comparison with untreated leaves in a choice test. Residence times of parasitoids on both leaves were recorded. The parasitoids seemed to prefer walking on leaves treated with homogenates of H. magnanima over untreated leaves, but the difference in residence times was not significant. In contrast, its residence time on leaves treated with homogenates of O. furnacalis was significantly shorter than that on untreated leaves. Thus, the induction of tea leaf surface chemicals may differ among moth species, which may produce different types of elicitors. This difference may, in turn, affect the host‐searching behaviour of A. reticulata.     

Effects of water stress on plant volatile emission and insect oviposition preference in an agroecosystem

1. Drought has become more common and severe in many parts of the world due to climate change. The effect of water stress on insect oviposition preference that is key determinant for their fitness has received less attention.
2. Here, we examined how water stress may affect oviposition selection of Ostrinia furnacalis for maize plants in the greenhouse, and tested difference in volatile compounds emitted from treated maize, and electronantennogram and bioassay responses of O. furnacalis to the volatile profiles in maize plants.
3. Ostrinia furnacalis were more prone to lay eggs on the well-watered maize. Most plant volatile compounds differed significantly among the three water treatments, including increased emissions of β-caryophyllene, (E)-2-hexenal, and linalool, and decreased emission of (Z)-3-hexen-1-ol when subjected to increasing intensity of drought.
4. Varied volatile profiles of maize may drive oviposition decision of O. furnacalis, because O. furnacalis showed a clear oviposition preference for (Z)-3-hexen-1-ol, while not for β-caryophyllene, (E)-2-hexenal, and linalool, at the concentration of 1000 ppm.
5. This study advances understanding of drought effects on plant–insect interactions through volatile profiles. Our finding calls for attention to oviposition selection for insect pest management in agricultural settings, especially in regions under changing precipitation patterns.

     

EFFECTS OF SUBLETHAL ESERINE ON THE CHEMICAL COMMUNICATION SYSTEM OF ASIAN CORN BORER,OSTRINIA FURNACALIS (GUENEE)*

Abstract Topically applied sublethal doses of eserine may interrupt chemical communication between the two sexes of Asian corn borer, Ostrinia furnacalis (Guenee), by affecting calling and sex pheromone titre released by the females. (1) Studies on the effects of sublethal eserine on the chemical communication system of O. furnacalis indicated that there was a decreasing probability of females calling and sex pheromone titre as the eserine dosage increased from 0.27 to 2 700 ng, with no effect on the periodicity. However, at 2 700 ng, the E/Z ratio of the sex pheromone components was affected. The sexual chemical communication system could not control within narrow level. (2) The recovery test of 27ng‐treated female indicated that the calling percentages of the 1st, 2nd and 4th day post‐treatment were 50%, 78% and 84% respectively. Sex pheromone titre was 40%, 40% and 80% of control female repectively. There was a trend toward recovery. The sexual chemical communication system could control within narrow level from the 2nd day post‐treatment.